Blood group, classification of blood based on inherited differences (polymorphisms) in antigens

on the surfaces of the red blood cells (erythrocytes). Inherited differences of white blood cells
(leukocytes), platelets (thrombocytes), and plasma proteins also constitute blood groups, but they
are not included in this discussion.

Historical Background

English physician William Harvey announced his observations on the circulation of the blood in 1616 and
published his famous monograph titled Exercitatio Anatomica de Motu Cordis et Sanguinis in Animalibus
(The Anatomical Exercises Concerning the Motion of the Heart and Blood in Animals) in 1628. His
discovery, that blood circulates around the body in a closed system, was an essential prerequisite of the
concept of transfusing blood from one animal to another of the same or different species. In England,
experiments on the transfusion of blood were pioneered in dogs in 1665 by physician Richard Lower. In
November 1667 Lower transfused the blood of a lamb into a man. Meanwhile, in France, Jean-Baptiste
Denis, court physician to King Louis XIV, had also been transfusing lambs’ blood into human subjects and
described what is probably the first recorded account of the signs and symptoms of a hemolytic
transfusion reaction. Denis was arrested after a fatality, and the procedure of transfusing the blood of
other animals into humans was prohibited, by an act of the Chamber of Deputies in 1668, unless
sanctioned by the Faculty of Medicine of Paris. Ten years later, in 1678, the British Parliament also
prohibited transfusions. Little advance was made in the next 150 years.

In England in the 19th century, interest was reawakened by the activities of obstetrician James
Blundell, whose humanitarian instincts had been aroused by the frequently fatal outcome of
hemorrhage occurring after childbirth. He insisted that it was better to use human blood for
transfusion in such cases. In 1875 German physiologist Leonard Landois showed that, if the red
blood cells of an animal belonging to one species are mixed with serum taken from an animal of
another species, the red cells usually clump and sometimes burst—i.e., hemolyze. He attributed
the appearance of black urine after transfusion of heterologous blood (blood from a different
species) to the hemolysis of the incompatible red cells. Thus, the dangers of transfusing blood of
another species to humans were established scientifically. The human ABO blood groups were
discovered by Austrian-born American biologist Karl Landsteiner in 1901. Landsteiner found
that there are substances in the blood, antigens and antibodies, that induce clumping of red cells
when red cells of one type are added to those of a second type. He recognized three groups—A,
B, and O—based on their reactions to each other. A fourth group, AB, was identified a year later
by another research team. Red cells of the A group clump with donor blood of the B group; those
of the B group clump with blood of the A group; those of the AB group clump with those of the
A or the B group because AB cells contain both A and B antigens; and those of the O group do
not generally clump with any group, because they do not contain either A or B antigens. The
application of knowledge of the ABO system in blood transfusion practice is of enormous
importance, since mistakes can have fatal consequences. The discovery of the Rh
system by Landsteiner and Alexander Wiener in 1940 was made
because they tested human red cells with antisera developed in rabbits
and guinea pigs by immunization of the animals with the red cells of
the rhesus monkey Macaca mulatta.
Other blood groups were identified later, such
as Kell, Diego, Lutheran, Duffy, and Kidd. The remaining blood group
systems were first described after antibodies were identified in
patients. Frequently, such discoveries resulted from the search for the
explanation of an unexpected unfavourable reaction in a recipient
after a transfusion with formerly compatible blood. In such cases the
antibodies in the recipient were produced against previously
unidentified antigens in the donor’s blood. In the case of the Rh
system, for example, the presence of antibodies in the maternal serum
directed against antigens present on the child’s red cells can have
serious consequences because of antigen-antibody reactions that
produce erythroblastosis fetalis, or hemolytic disease of the newborn.
Some of the other blood group systems—for example, the Kell and
Kidd systems—were discovered because an infant was found to have
erythroblastosis fetalis even though mother and child were compatible
as far as the Rh system was concerned. In the table the well-
established human blood group systems are listed in the order of
discovery.

Major human blood group systems

system date of discovery main antigens

ABO 1901 A1, A2, B, H

MNSs 1927 M, N, S, s

P 1927 P1, P2

Rh 1940 D, C, c, E, e

Lutheran 1945 Lua, Lub

Kell 1946 K, k

Lewis 1946 Lea, Leb

 Major human blood group systems

system date of discovery main antigens

Duffy 1950 Fya, Fyb

Kidd 1951 Jka, Jkb

Diego 1955 Dia, Dib

Yt 1956 Yta, Ytb

I 1956 I, i

Xg 1962 Xga

Dombroc
1965 Doa
k

The Importance Of Antigens And Antibodies

The red cells of an individual contain antigens on their surfaces that correspond to their blood group and
antibodies in the serum that identify and combine with the antigen sites on the surfaces of red cells of
another type. The reaction between red cells and corresponding antibodies usually results in clumping—
agglutination—of the red cells; therefore, antigens on the surfaces of these red cells are often referred
to as agglutinogens. Antibodies are part of the circulating plasma proteins known as immunoglobulins,
which are classified by molecular size and weight and by several other biochemical properties. Most
blood group antibodies are found either on immunoglobulin G (IgG) or immunoglobulin M (IgM)
molecules, but occasionally the immunoglobulin A (IgA) class may exhibit blood group specificity.
Naturally occurring antibodies are the result of immunization by substances in nature that have
structures similar to human blood groups. These antibodies are present in an individual despite the fact
that there has been no previous exposure to the corresponding red cell antigens—for example, anti-A in
the plasma of people of blood group B and anti-B in the plasma of people of blood group A.  Immune
antibodies are evoked by exposure to the corresponding red cell antigen. Immunization (i.e., the
production of antibodies in response to antigen) against blood group antigens in humans can occur as a
result of pregnancy, blood transfusion, or deliberate immunization. The combination of pregnancy and
transfusion is a particularly potent stimulus. Individual blood group antigens vary in their antigenic
potential; for example, some of the antigens belonging to the Rh and ABO systems are strongly
immunogenic (i.e., capable of inducing antibody formation), whereas the antigens of the Kidd and Duffy
blood group systems are much weaker immunogens.

The blood group antigens are not restricted solely to red cells or even to hematopoietic tissues. The
antigens of the ABO system are widely distributed throughout the tissues and have been unequivocally
identified on platelets and white cells (both lymphocytes and polymorphonuclear leukocytes) and in
skin, the epithelial (lining) cells of the gastrointestinal tract, the kidney, the urinary tract, and the lining
of the blood vessels. Evidence for the presence of the antigens of other blood group systems on cells
other than red cells is less well substantiated. Among the red cell antigens, only those of the ABO system
are regarded as tissue antigens and therefore need to be considered in organ transplantation.

Chemistry Of The Blood Group Substances

The exact chemical structure of some blood groups has been identified, as have the gene products (i.e.,
those molecules synthesized as a result of an inherited genetic code on a gene of a chromosome) that
assist in synthesizing the antigens on the red cell surface that determine the blood type. Blood
group antigens are present on glycolipid and glycoprotein molecules of the red cell membrane.
The carbohydrate chains of the membrane glycolipids are oriented toward the external surface of the
red cell membrane and carry antigens of the ABO, Hh, Ii, and P systems. Glycoproteins,
which traverse the red cell membrane, have a polypeptide backbone to which carbohydrates are
attached. An abundant glycoprotein, band 3, contains ABO, Hh, and Ii antigens.
Another integral membrane glycoprotein, glycophorin A, contains large numbers of sialic acid molecules
and MN blood group structures; another, glycophorin B, contains Ss and U antigens.

The genes responsible for inheritance of ABH and Lewis antigens are glycosyltransferases (a group of
enzymes that catalyze the addition of specific sugar residues to the core precursor substance). For
example, the H gene codes for the production of a specific glycosyltransferase that adds L-fucose to a
core precursor substance, resulting in the H antigen; the Le gene codes for the production of a specific
glycosyltransferase that adds L-fucose to the same core precursor substance, but in a different place,
forming the Lewis antigen; the A gene adds N-acetyl-D-galactosamine (H must be present), forming the
A antigen; and the B gene adds D-galactose (H must be present), forming the B antigen. The P system
is analogous to the ABH and Lewis blood groups in the sense that the P antigens are built by the addition
of sugars to precursor globoside and paragloboside glycolipids, and the genes responsible for these
antigens must produce glycosyltransferase enzymes.

The genes that code for MNSs glycoproteins change two amino acids in the sequence of the
glycoprotein to account for different antigen specificities. Additional analysis of red cell membrane
glycoproteins has shown that in some cases the absence of blood group antigens is associated with an
absence of minor membrane glycoproteins that are present normally in antigen-positive persons.
Methods Of Blood Grouping

Identification of blood groups

The basic technique in identification of the antigens and antibodies of blood groups is the agglutination
test. Agglutination of red cells results from antibody cross-linkages established when different specific
combining sites of one antibody react with antigen on two different red cells. By mixing red cells
(antigen) and serum (antibody), either the type of antigen or the type of antibody can be determined
depending on whether a cell of known antigen composition or a serum with known antibody specificity
is used.

In its simplest form, a volume of serum containing antibody is added to a thin suspension (2–5 percent)
of red cells suspended in physiological saline solution in a small tube with a narrow diameter. After
incubation at the appropriate temperature, the red cells will have settled to the bottom of the tube.
These sedimented red cells are examined macroscopically (with the naked eye) for agglutination, or they
may be spread on a slide and viewed through a low-power microscope.

An antibody that agglutinates red cells when they are suspended in saline solution is called a complete
antibody. With powerful complete antibodies, such as anti-A and anti-B, agglutination reactions visible
to the naked eye take place when a drop of antibody is placed on a slide together with a drop containing
red cells in suspension. After stirring, the slide is rocked, and agglutination is visible in a few minutes. It
is always necessary in blood grouping to include a positive and a negative control for each test.

An antibody that does not clump red cells when they are suspended in saline solution is
called incomplete. Such antibodies block the antigenic sites of the red cells so that subsequent addition
of complete antibody of the same antigenic specificity does not result in agglutination. Incomplete
antibodies will agglutinate red cells carrying the appropriate antigen, however, when the cells are
suspended in media containing protein. Serum albumin from the blood of cattle is a substance that is
frequently used for this purpose. Red cells may also be rendered specifically agglutinable by incomplete
antibodies after treatment with such protease enzymes as trypsin, papain, ficin, or bromelain.

After such infections as pneumonia, red cells may become agglutinable by almost all normal sera
because of exposure of a hidden antigenic site (T) as a result of the action of bacterial enzymes. When
the patient recovers, the blood also returns to normal with respect to agglutination. It is unusual for the
red cells to reflect antigenicity other than that determined by the individual’s genetic makeup. The
presence of an acquired B antigen on the red cells has been described occasionally in diseases of the
colon, thus allowing the red cell to express an antigenicity other than that genetically determined. Other
diseases may alter immunoglobulins; for example, some may induce the production of antibodies
directed against the person’s own blood groups (autoimmune hemolytic anemia) and thus may interfere
with blood grouping. In other diseases a defect in antibody synthesis may cause the absence of anti-A
and anti-B antibody.

Coombs test

When an incomplete antibody reacts with the red cells in saline solution, the antigenic sites become
coated with antibody globulin (gamma globulin), and no visible agglutination reaction takes place. The
presence of gamma globulin on cells can be detected by the Coombs test, named for its inventor, English
immunologist Robert Coombs. Coombs serum (also called antihuman globulin) is made by immunizing
rabbits with human gamma globulin. The rabbits respond by making antihuman globulin (i.e., antibodies
against human gamma globulin and complement) that is then purified before use. The antihuman
globulin usually contains antibodies against IgG and complement. Coombs serum is added to the
washed cells; the tube is centrifuged; and, if the cells are coated by gamma globulin or complement,
agglutinates will form. Newer antiglobulin reagents (made by immunizing with purified protein) can
detect either globulin or complement. Depending on how it is performed, the Coombs test can detect
incomplete antibody in the serum or antibody bound to the red cell membrane. In certain
diseases, anemia may be caused by the coating of red cells with gamma globulin. This can happen when
a mother has made antibodies against the red cells of her newborn child or if a person makes an
autoantibody against his own red cells.

Adsorption, elution, and titration

If a serum contains a mixture of antibodies, it is possible to prepare

pure samples of each by a technique called adsorption. In this
technique an unwanted antibody is removed by mixing it with red cells
carrying the appropriate antigen. The antigen interacts with the
antibody and binds it to the cell surface. These red cells are washed
thoroughly and spun down tightly by centrifugation, all the fluid above
the cells is removed, and the cells are then said to be packed. The cells
are packed to avoid dilution of the antibody being prepared.
Adsorption, then, is a method of separating mixtures of antibodies by
removing some and leaving others. It is used to identify antibody
mixtures and to purify reagents. The purification of the Coombs serum
(see above) is done in the same way.

If red cells have adsorbed gamma globulin onto their surfaces, the
antibody can sometimes be recovered by a process known as elution.
One simple way of eluting (dissociating) antibody from washed red
cells is to heat them at 56 °C (133 °F) in a small volume of saline
solution. Other methods include use of acid or ether. This technique is
sometimes useful in the identification of antibodies.

Titration is used to determine the strength of an antibody. Doubling

dilutions of the antibody are made in a suitable medium in a series of
tubes. Cells carrying the appropriate antigen are added, and the
agglutination reactions are read and scored for the degree of positivity.
The actual concentration of the antibody is given by the dilution at
which some degree of agglutination, however weak, can still be seen.
This would not be a safe dilution to use for blood-grouping purposes.
If an antiserum can be diluted, the dilution chosen must be such that
strong positive reactions occur with selected positive control cells.
Titration is helpful when preparing reagents and comparing antibody
concentrations at different time intervals.

Inhibition tests

Inhibition tests are used to detect the presence of antigen

with blood group specificity in solutions; inhibition of a known
antibody-antigen reaction by a fluid indicates a particular blood group
specificity. If an active substance is added to antibody, neutralization
of the antibody’s activity prevents agglutination when red cells
carrying the appropriate antigen are subsequently added to the
mixture. A, B, Lewis, Chido, Rogers, and P antigens are readily
available and can be used to facilitate antibody identification. This
technique was used to elucidate the biochemistry of ABH, Ii, and
Lewis systems, and it is important in forensic medicine as a means of
identifying antigens in blood stains.

Hemolysis

Laboratory tests in which hemolysis (destruction) of the red cells is the

end point are not used frequently in blood grouping. For hemolysis to
take place, a particular component of fresh serum
called complement must be present. Complement must be added to
the mixture of antibody and red cells. It may sometimes be desirable
to look for hemolysins that destroy group A red cells in mothers whose
group A children are incompatible or in individuals, not belonging to
groups A or AB, who have been immunized with tetanus toxoid that
contains substances with group A specificity.

Hemolytic reactions may occur in patients who have been

given transfusions of blood that either is incompatible or has already
hemolyzed. The sera of such patients require special investigations to
detect the presence of hemoglobin that has escaped from red cells
destroyed within the body and for the breakdown products of other
red cell constituents.

Sources of antibodies and antigens

Normal donors are used as the source of supply of naturally occurring

antibodies, such as those of the ABO, P, and Lewis systems. These
antibodies work best at temperatures below that of the body (37 °C, or
98.6 °F); in the case of what are known as cold agglutinins, such as
anti-P1, the antibody is most active at 4 °C (39 °F). Most antibodies
used in blood grouping must be searched for in immunized donors.

Antibodies for MN typing are usually raised in rabbits—similarly for

the Coombs serum. Antibodies prepared in this way have to be
absorbed free of unwanted components and carefully standardized
before use. Additional substances with specific blood group activity
have been found in certain plants. Plant agglutinins are called lectins.
Some useful reagents extracted from seeds are anti-H from Ulex
europaeus (common gorse); anti-A1, from another member of the
pulse family Fabaceae (Leguminosae), Dolichos biflorus; and anti-N
from the South American plant Vicia graminea. Agglutinins have also
been found in animals—for example, the fluid pressed from the land
snail Octala lactea. Additional plant lectins and agglutinins from
animal fluids have been isolated.

Monoclonal antibodies (structurally identical antibodies produced by

hybridomas) to blood groups are replacing some of the human blood
grouping reagents. Mouse hybridomas (hybrid cells of a myeloma
tumour cell and lymphocyte merging) produce anti-A and anti-B
monoclonal antibodies. The antibodies are made by immunizing with
either red cells or synthetic carbohydrates. In addition to their use in
blood grouping, these monoclonal antibodies can be of use in defining
the hereditary background (heterogenicity) and structure of the red
cell antigen.
Uses Of Blood Grouping

Transfusion

The blood donated by healthy persons is tested to ensure that the level
of hemoglobin is satisfactory and that there is no risk of transmitting
certain diseases, such as AIDS or hepatitis. It is then fractionated
(split) into its component parts, particularly red cells, plasma, and
platelets. Correct matching for the ABO system is vital. Compatible
donors on the basis of their possessing A, B, or O blood are shown in
the table.

The ABO and Rh groups in transfusion

system recipient type donor red cell type donor plasma type

*Not if the patient's serum contains anti-A1 (antibody to common type A red cell in subgroup A patients).

**Not if the patient is a female less than 45 years old (childbearing possible), unless life-threatening hemorrhage is
present and transfusion of Rh-positive blood is lifesaving.

***Not if the patient's serum contains anti-D (antibody to positive red cells), except under unusual medical
circumstances.

ABO A A* or O A or AB

ABO B B or O B or AB

ABO O O only O, A, B, or AB

ABO AB AB*, A*, B, or O AB

Rh positive positive or negative positive or negative

The ABO and Rh groups in transfusion

system recipient type donor red cell type donor plasma type

Rh negative negative or positive**, *** negative or positive**

As explained above, the most important blood group systems for

transfusion of red cells are ABO and Rh. Persons who have either of
the red cell antigens (A and B) have antibody present in their serum of
the type that will oppose an antigen of its opposite nature; for
example, group A blood contains A antigens on red cell surfaces and
anti-B antibodies in the surrounding serum. On the other hand, O
group individuals lack both the A and the B antigen and thus have
both anti-A and anti-B in their serum. If these antibodies combine
with the appropriate antigen, the result is hemolytic transfusion
reaction and possibly death. Red cell transfusions must therefore be
ABO compatible. The blood groups A and B have various subgroups
(e.g., A1, A2, A3, and B1, B2, and B3). The only common subgroups that
are likely to affect red cell transfusions are the subgroups of A.

Potential donors are also tested for some of the antigens of the Rh
system, since it is essential to know whether they are Rh-positive or
Rh-negative. Rh-negative indicates the absence of the D antigen. Rh-
negative persons transfused with Rh-positive blood will make anti-D
antibodies from 50 to 75 percent of the time. Antibody made in
response to a foreign red cell antigen is usually not harmful but does
require subsequent transfusions to be antigen-negative. Rh-positive
blood should never be given to Rh-negative females before or during
the childbearing age unless Rh negative blood is not available and the
transfusion is lifesaving. If such a woman subsequently
became pregnant with an Rh-positive fetus, she might form anti-Rh
antibody, even though the pregnancy was the first, and the child might
develop erythroblastosis fetalis (hemolytic disease of the newborn).

Care must be taken not to give a transfusion unless the cells of the
donor have been tested against the recipient’s serum. If this
compatibility test indicates the presence of antibodies in the
recipient’s serum for the antigens carried by the donor’s cells, the
blood is not suitable for transfusion because an unfavourable reaction
might occur. The test for compatibility is called the direct match test.
It involves testing the recipient’s serum with the donor’s cells and by
the indirect Coombs test. These are adequate screening tests for most
naturally occurring and immune antibodies.

If, in spite of all the compatibility tests, a reaction does occur after the
transfusion is given (the unfavourable reaction often manifests itself in
the form of a fever), an even more careful search must be made for any
red cell antibody that might be the cause. A reaction after transfusion
is not necessarily due to red cell antigen-antibody reactions. It could
be caused by the presence of antibodies to the donor’s platelets or
white cells. Transfusion reactions are a particular hazard for persons
requiring multiple transfusions.

Organ transplants

The ABO antigens are widely distributed throughout the tissues of the
body. Therefore, when organs such as kidneys are transplanted, most
surgeons prefer to use organs that are matched to the recipient’s with
respect to the ABO antigen system, although the occasional survival of
a grafted ABO-incompatible kidney has occurred. The remaining red
cell antigen systems are not relevant in organ transplantation.

Paternity testing

Although blood group studies cannot be used to prove paternity, they

can provide unequivocal evidence that a male is not the father of a
particular child. Since the red cell antigens are inherited as dominant
traits, a child cannot have a blood group antigen that is not present in
one or both parents. For example, if the child in question belongs to
group A and both the mother and the putative father are group O, the
man is excluded from paternity. The table shows the phenotypes
(observed characters) of the offspring that can and cannot be
produced in the matings on the ABO system, considering only the
three alleles (alternative genes) A, B, and O. Similar inheritance
patterns are seen in all blood group systems. Furthermore, if one
parent is genetically homozygous for a particular antigen—that is, has
inherited the gene for it from both the grandfather and grandmother
of the child—then that antigen must appear in the blood of the child.
For example, on the MN system, a father whose phenotype is M and
whose genotype is MM (in other words, a man who is of blood type M
and has inherited the characteristic from both parents) will transmit
an M allele to all his progeny.

Exclusions of paternity on the ABO system

possible
matings impossible children
children

O×O O A, B, AB

O×A O, A B, AB

O×B O, B A, AB

O × AB A, B O, AB

A×A O, A B, AB

A×B O, A, B, AB

A × AB A, B, AB O

B×B O, B A, AB
Exclusions of paternity on the ABO system

possible
matings impossible children
children

B × AB A, B, AB O

AB × AB A, B, AB O

In medicolegal work it is important that the blood samples are

properly identified. By using multiple red cell antigen systems and
adding additional studies on other blood types (HLA
[human leukocyte antigen], red cell enzymes, and plasma proteins), it
is possible to state with a high degree of statistical certainty that a
particular male is the father.

Blood Groups And Disease

In some cases an increased incidence of a particular antigen seems to

be associated with a certain disease. Stomach cancer is more common
in people of group A than in those of groups O and B. Duodenal
ulceration is more common in nonsecretors of ABH substances than in
secretors. For practical purposes, however, these statistical
correlations are unimportant. There are other examples that illustrate
the importance of blood groups to the normal functions of red cells.

In persons who lack all Rh antigens, red cells of altered shape

(stomatocytes) and a mild compensated hemolytic anemia are present.
The McLeod phenotype (weak Kell antigens and no Kx antigen) is
associated with acanthocytosis (a condition in which red cells have
thorny projections) and a compensated hemolytic anemia. There is
evidence that Duffy-negative human red cells are resistant to infection
by Plasmodium knowlesi, a simian malaria parasite. Other studies
indicate that P. falciparum receptors may reside on glycophorin A and
may be related to the Wrb antigen.

Blood group incompatibility between mother and child can

cause erythroblastosis fetalis (hemolytic disease of the newborn). In
this disease IgG blood group antibody molecules cross the placenta,
enter the fetal circulation, react with the fetal red cells, and destroy
them. Only certain blood group systems cause erythroblastosis fetalis,
and the severity of the disease in the fetus varies greatly. ABO
incompatibility usually leads to mild disease. Rh, or D antigen,
incompatibility is now largely preventable by treating Rh-negative
mothers with Rh immunoglobulin, which prevents immunization
(forming antibodies) to the D antigen. Many other Rh antigens, as well
as other red cell group antigens, cause erythroblastosis fetalis. The
baby may be anemic at birth, which can be treated by transfusion with
antigen-negative red cells. Even total exchange transfusion may be
necessary. In some cases, transfusions may be given while the fetus is
still within the uterus (intrauterine transfusion). Hyperbilirubinemia
(an increased amount of bilirubin, a breakdown product
of hemoglobin, in the blood) may lead to neurological deficits.
Exchange transfusion eliminates most of the hemolysis by providing
red cells, which do not react with the antibody. It also decreases the
amount of antibody and allows the child to recover from the disease.
Once the antibody disappears, the child’s own red cells survive
normally.

Genetic And Evolutionary Significance Of

Blood Groups
Blood groups and genetic linkage

Red cell groups act as markers (inherited characteristics)

for genes present on chromosomes, which are responsible for their
expression. The site of a particular genetic system on a chromosome is
called a locus. Each locus may be the site of several alleles (alternative
genes). In an ordinary cell of the human body, there are 46
chromosomes arranged in 23 pairs, 22 pairs of which are autosomes
(chromosomes other than sex chromosomes), with the remaining pair
being the sex chromosomes, designated XX in females and XY in
males. The loci of the blood group systems are on the autosomes,
except for Xg, which is unique among the blood groups in being
located on the X chromosome. Genes carried by the X chromosome
are said to be sex-linked. Since the blood groups are inherited in a
regular fashion, they can be used as genetic markers in family studies
to investigate whether any two particular loci are sited on the same
chromosome—i.e., are linked. The genes sited at loci on the same
chromosome travel together from parent to child, and, if the loci are
close together, the genes will rarely be separated.

Loci that are farther apart can be separated by recombination. This

happens when material is exchanged between homologous
chromosomes (pair of chromosomes) by crossing over during the
process of cell division (mitosis). The reproductive cells contain half
the number of chromosomes of the rest of the body; ova carry an X
chromosome and spermatozoa an X or a Y. The characteristic number
of 46 chromosomes is restored at fertilization. In a classical pedigree
linkage study, all the members of a family are examined for a test
character and for evidence of the nonindependent segregation of pairs
of characters. The results must be assessed statistically to determine
linkage. Individual chromosomes are identified by the banding
patterns revealed by different staining techniques. Segments of
chromosomes or chromosomes that are aberrant in number
and morphology may be precisely identified. Other methods for
localizing markers on chromosomes include somatic cell hybridization
(cell culture with alignment of single strands of RNA and DNA) and
use of DNA probes (strands of radiolabeled DNA). These methods are
useful in classical linkage studies to locate blood group loci. The loci
for many red cell groups have been found on chromosomes and in
many cases have been further localized on a particular chromosome.

In some of the blood group systems, the amount of antigen produced

depends on the genetic constitution. The ABO blood group gene codes
for a specific carbohydrate transferase enzyme that catalyzes the
addition of specific sugars onto a precursor substance. As a new sugar
is added, a new antigen is produced. Antigens in the MNSs blood
system are the products of genes that control terminal amino
acid sequence. The amount of antigen present may depend on the
amount of gene product inherited or on the activity of the gene
product (i.e., transferase). The red cells of a person
whose genotype is MM show more M antigen than do MN red cells. In
the case of ABO, the same mechanism may also play a role in antigen
expression, but specific activity of the inherited transferase may be
more important.

The amount of antigen produced can also be influenced by the

position of the genes. Such effects within a genetic complex can be due
to determinants on the same chromosome—they are then said to be cis
—or to determinants on the opposite chromosome of a chromosome
pair—trans.

In the Rh combination cdE/cde, more E antigen is produced than in

the combination cDE/cde. This may be due to the suppressor effect of
D on E. An example of suppression in the trans situation is that more
C antigen is detectable on the red cells from CDe/cde donors than on
those of CDe/cDE people. The inheritance of the Rh system probably
depends on the existence of operator genes, which turn the activity of
closely linked structural genes on or off. The operator genes are
themselves controlled by regulator genes. The operator genes are
responsible for the quantity of Rh antigens, while the structural genes
are responsible for their qualitative characteristics.

The detection of recombination (exchange of material between

chromosomes) or mutation in human families is complicated by
questions of paternity. In spite of the large number of families that
have been studied, it is an extremely rare occurrence. The paucity of
examples may indicate that the recombinant and mutation rate for
blood group genes is lower than that estimated for other human genes.

Blood groups and population groups

The blood groups are found in all human populations but vary in
frequency. An analysis of populations yields striking differences in the
frequency of some blood group genes. The frequency of the A gene is
the highest among Australian Aborigines, the Blackfoot Indians of
Montana in the United States, and the Sami people of northern
Scandinavia. The O gene is common throughout the world,
particularly among peoples of South and Central America. The
maximum frequency of the B gene occurs in Central Asia and northern
India. On the Rh system most northern and central European
populations differ from each other only slightly and are characterized
by a cde (r) frequency of about 40 percent. Africans show a
preponderance of the complex cDe, and the frequency of cde is about
20 percent. In eastern Asia cde is almost wholly absent, and, since
everyone has the D antigen, erythroblastosis fetalis (due to the
presence of maternal anti-D) is unknown in these populations.

The blood group frequencies in small inbred populations reflect the

influences of genetic drift. In a small community an allele can be lost
from the genetic pool if persons carrying it happen to be infertile,
while it can increase in frequency if advantage exists. It has been
suggested, for example, that B alleles were lost by chance from Native
Americans and Australian Aborigines when these communities were
small. There are pronounced discrepancies in blood group frequencies
between the people of eastern Asia and the aboriginal peoples of the
Americas. Other blood group frequencies in different populations
show that ancestors might share some common attribute indicating a
close resemblance between populations.

Nonhuman primates carry blood group antigens that can be detected

with reagents used for typing human beings. The closer their
evolutionary relationship to humans, the greater their similarity with
respect to antigens. The red cells of the apes, with the exception of
the gorilla, have ABO antigens that are indistinguishable from those of
human cells. Chimpanzees and orangutans are most frequently group
A, but groups O, B, and AB are represented. Gibbons can be of any
group except O, and gorillas have a B-like antigen that is not identical
in activity with the human one. In both Old and New World monkeys,
the red cells do not react with anti-A or with anti-B, but, when the
secretions are examined, A and B substances and agglutinins are
present in the serum. As far as the Rh system is concerned,
chimpanzees carry two Rh antigens—D and c (hr′)—but not the others,
whereas gibbons have only c (hr′). The red cells of monkeys do not
give clear-cut reactions with human anti-Rh sera.